Glycolysis: Energy

Generation Without
an Oxygen
Requirement

Glucose Biofuel Prominence:

Low-reactive ring-form
minimizes protein glycosylation
Glycolysis: A Three
Step Process
• Glucose trapping and
destabilization (priming)

•Three carbon unit generation

(cleaving)

• Energy generation
Induced Fit in
Hexokinase
Glucose induces a
large enzyme
conformational
change

Substrate-induced cleft
closing prevents ATP hydrolysis

Kinases require a divalent metal ion

What function does Mg+2 play in hexokinase?

Hexokinase Closed Around Substrates
What
mechanisms
of catalysis
are
operative?
Hexokinase Reaction Mechanism

What is the Nu:, electrophile, and leaving

group in this reaction?
Phosphoglucose
Isomerase:
Aldose to
Ketose
Conversion
Phosphoglucose Isomerase (PGI) G6P
Conversion via Acid-Base Catalysis

PGI Reaction Mechanism

Phosphoglucose Isomerase (PGI) G6P
Conversion via Acid-Base Catalysis

Base catalyzed
bond formation
Phosphoglucose Isomerase (PGI) G6P
Conversion via Acid-Base Catalysis

Acid catalyzed
ketal formation
Phosphoglucose Isomerase (PGI) G6P
Conversion via Acid-Base Catalysis
Base catalyzes ring closure

H+
Phosphofructokinase: Trapping the
Fructose Isomer

What is the mechanism for this reaction?

Glycolysis Stage I: Glucose Trapping
and Destabilization (priming)
Six Carbon Sugar Cleaved to Two
Three Carbon Units

What is the bond to be cleaved?

Which alcohol becomes an aldehyde?
Haworth and Fischer Projections
Equivalency

The functional group that is down in a Haworth

projection is positioned how in a Fischer
structure?
 Aldolase Reaction Mechanism

Aldolase

Fructose-1,6-bisphosphate binds to the aldolase

enzyme for covalent catalysis
 Aldolase Reaction Mechanism

What is lost when the Schiff base forms?

Aldolase Reaction Mechanism

Aldolase Rxn Mechanism

Compare and contrast a Schiff base with a

carbonyl group.
Aldolase Reaction Mechanism

What is the process for Schiff base to carbonyl

conversion?
Aldolase Reaction Mechanism

H2O

Aldolase cleaves FBP into GAP and DHAP

Triose Phosphate Isomerase (TIM)
Reversible and driven towards GAP due to product depletion

Which previous glycolytic step is similar to TIM?

Triose Phosphate Isomerase
Reaction Mechanism

Glycolysis: Step #5
Triose Phosphate Isomerase
TIM- or α,β-barrel
with 8 parallel β-
strands surrounded
by 8 α-helices.

DHAP conversion to GAP

necessary to proceed
through glycolysis
Stoichiometry: Stages 1-2 of Glycolysis

Two ATPs are initially

invested.

One glucose is
metabolized into two
GAP molecules.
Road
Map for
Energy
Harvest
(Stage 3)
Glyceraldehyde-3-Phosphate
Dehydrogenase: Covalent Catalysis
Glyceraldehyde-3-Phosphate
Dehydrogenase: a 2 Step Process

What amino acid will serve as a nucleophile to

form a thioester?
Glyceraldehyde-3-Phosphate
Dehydrogenase:
Reaction
Mechanism
Glyceraldehyde-3-Phosphate
Dehydrogenase: Catalysis Energetics
Hypothetical reaction with no coupling

Actual coupled reaction

Phosphoglycerate Kinase

What is the Nu:, electrophile and leaving group for

this reaction? (hint: consider hexokinase in
reverse)
Glycolysis: the Three Final Steps
Pyruvate Kinase

What is the Nu:, electrophile and leaving group for

this reaction? (hint: consider phosphoglycerate
kinase)
Glycolysis Energetic
∆G°ʹ ∆G
Enzyme (kcal/mol) (kcal/mol)

1near equilibrium means that ∆G is about zero

What is the relationship between ∆G and ∆G°ʹ?
When can ∆G and ∆G°ʹ diverge?
Regulating Glycolysis: A Pictorial Analogue
• Water represents
metabolite flux

• Water amount in
flask represents
intermediate
abundance

• Flasks connections
are enzymes

• Vertical drop represents

decrease in free energy

ΔG° = height difference between flask bottoms

ΔG = height difference between water levels
Metabolic Regulation

Irreversible reactions are potential regulatory sites

(e.g. hexokinase, phosphofructokinase and pyruvate
kinase)

What duel role does

ATP play in PFK-1
catalysis?

In what direction does

ATP regulate
phosphofructokinase?
Energy Status Regulates Glycolytic Flow
Elevated [ATP] sufficient energy; elevate [AMP] low energy
ADP + ADP ↔ ATP + AMP <adenylate kinase>
Muscle Tissue
Fructose-2,6-Bisphosphate an Allosteric
Regulator of Phosphofructokinase-1
Liver Tissue

PFK-2

Front activation by fructose-6P

F-2,6-BP amplifies or diminishes PFK-1 activity?
Fructose-2,6-Bisphosphate Reduces ATP
Inhibition of Phosphofructokinase-1

Liver Tissue

PFK-2

ATP is a substrate and

inhibitor of PFK-1
Fructose Entry Points for
Glycolysis

Glucose + Fructose Glycerol-3P

Major dietary sugars: sucrose (table sugar) and

fructose (high-fructose corn syrup)
Fructose Metabolism

How is
this
different
than
glucose
metabolism?
Fructose Metabolism
Glycerol 3-phosphate a precursor to triacylglycerol

Fructose catabolism bypasses

phosphofructokinase regulation

Lipid Synthesis
Alternative Fates for Pyruvate
Anaerobic Recycling of
NADH for Glycolysis
Microbial Recycling of
NADH for Glycolysis
 Pyruvate
Dehydrogenase:
the Bridge
between
Glycolysis and
Citric Acid Cycle
Standard Free Energy Change
Comparisons for Glucose Catabolism
With and Without Oxygen
Pathogenic Obligate Anaerobes
Pyruvate Targeted for Anabolism

The biotin
prosthetic
group serves as
a CO2 carrier

What reaction
links biotin to the
protein?
 Pyruvate
Carboxylase: an
Endergonic
Reaction

Oxaloacetate
Glucose Metabolism: Both Catabolic and
Anabolic
Glucose Metabolism: Both Catabolic and
Anabolic
Problems: 1, 3, 5, 7, 13, and 21